Hi there,
As an approach to characterizing the phosphorylation pattern of a nuclear
protein, I'd like to separate its different phospho-forms by IEF. For
starters, I'd be happy to see a couple of focussed bands, either from a
cell extract or an immunoprecipitate. The theoretical IP (w/o phosphates)
is 5.9, and the thing is kinda large (100kD). Does anybody out there have
some experience with a similar problem? Any particular pitfalls to look
for? Hints are greatly appreciated!
Oh, yes. I can use either vertical slab systems or a flatbed apparatus
(Pharmacia Multiphor II).
thanx in advance,
roney